College of Pharmacy and Health Sciences, Butler University, 4600 Sunset Avenue, Indianapolis, IN 46208, USA.
Nucleic Acids Res. 2010 Mar;38(5):1441-9. doi: 10.1093/nar/gkp1130. Epub 2009 Dec 16.
Chromatin remodeling is an essential part of transcription initiation. We show that at heat shock gene promoters functional interactions between individual ATP-dependent chromatin remodeling complexes play critical role in both nucleosome displacement and Pol II recruitment. Using HSP12, HSP82 and SSA4 gene promoters as reporters, we demonstrated that while inactivation of SNF2, a critical ATPase of the SWI/SNF complex, primarily affects the HSP12 promoter, depletion of STH1- a SNF2 homolog from the RSC complex reduces histone displacement and abolishes the Pol II recruitment at all three promoters. From these results, we conclude that redundancy between SWI/SNF and RSC complexes is only partial and likely is affecting different chromatin remodeling steps. While inactivation of other individual ATP-dependent chromatin remodeling complexes negligibly affects reporter promoters, combinatorial inactivation of SNF2 and ISW1 has a synergistic effect by diminishing histone loss during heat induction and eliminating Pol II recruitment. Importantly, it also eliminates preloading of HSF on HSP82 and SSA4 promoters before heat shock and diminishes HSF binding during heat shock. These observations suggest that prior action of chromatin remodeling complexes is necessary for the activator binding.
染色质重塑是转录起始的一个重要组成部分。我们表明,在热休克基因启动子中,单个 ATP 依赖的染色质重塑复合物之间的功能相互作用在核小体位移和 Pol II 募集中都起着关键作用。使用 HSP12、HSP82 和 SSA4 基因启动子作为报告基因,我们证明,虽然 SWI/SNF 复合物的关键 ATP 酶 SNF2 的失活主要影响 HSP12 启动子,但 RSC 复合物中的 STH1(SNF2 的同源物)的消耗减少了组蛋白位移,并取消了所有三个启动子的 Pol II 募集。根据这些结果,我们得出结论,SWI/SNF 和 RSC 复合物之间的冗余只是部分的,可能影响不同的染色质重塑步骤。虽然其他单个 ATP 依赖的染色质重塑复合物的失活对报告基因启动子的影响可以忽略不计,但 SNF2 和 ISW1 的组合失活通过减少热诱导过程中组蛋白的丢失并消除 Pol II 的募集,具有协同作用。重要的是,它还消除了 HSF 在热休克前在 HSP82 和 SSA4 启动子上的预加载,并在热休克期间减少了 HSF 的结合。这些观察结果表明,染色质重塑复合物的先前作用对于激活剂结合是必要的。